1, 11-iminoestratetraenes



United States Patent This invention relates to new steroid compounds.More particularly, it relates to 1,1 1-iminoestra-1,3 ,5 ,9(1 1tetraenes and their preparation.

The novel steroids of this invention can be illustrated by the followingformula:

wherein R is selected from the group consisting of hydrogen and loweralkyl; R is selected from the group consisting of hydrogen, lower alkyl,lower alkalnoyl, mono-nuclear aroyl, and lower alkoxy mono-nucleararoyl; X is selected from the group consisting of H OH OH OH 0:0; 0 O;/G and /C OH doweralkyl onon, 05011 and acid addition and quaternaryammonium salts thereof.

The present compounds are, in general, crystalline solids relativelyinsoluble in organic solvents such as lower alkyl alcohols, acetone,ethyl acetate, benzene, toluene, chloroform, ether, petroleum ether andthe like.

The present compounds which are described broadly as1,11-iminoestra-1,3,5 (10),9(11)-tetraenes substituted in the17-position can be made from the appropriate 1,11-iminoestratn'enes byheating with palladium-on-carbon catalyst. The1,11-iminoestra-l,3,5(l0),9(l1)-tetraenes are obtained from thecorresponding 1,11iminoestra-1,3,5(10)-trienes described and claimed inour copending application Serial No. 362,185, filed April 23, 1964 byheating the latter with palladium-on-carbon catalyst in the presence ofa solvent.

Using the procedure described hereinafter among the 4-amino-derivatives,which may be used as starting materials, are the following:4-amino-3-methoxy-l9-norpregna-1,3,5(10)-trien-20-one; 4-amino 116acetoxy-3- methoxyestra-l,3,5 10)-trien-17-one; 4-arnino l7,20;20,2l-bismethylenedioxy-3-methoxy 19 norpregna 1,3,5 (10)-trien-11fi-ol 11acetate; 4 amino-3-methoxyestra- 1,3,5(10)-trien-l6a,17}3-diol16,17-diacetate and 4-amino- 3-methoxyestra-1,3,5 10)-trien-17B-ol-16-one 17 -acetate.

The above compounds are prepared from the corresponding compoundslacking a 4-amino substituent by nitration (nitric acidacetic acid) andmethylation (dimethylsulfate), followed by reduction (sodiumhydrosulfite), e.g., estrone- 4 nitroestrone 4 nitroestrone methyl ether4-aminoestrone methyl ether.

The 4-amino steroids described above may be transformed into thefollowing l-amino compounds by methods described hereinafter:l-amino-4-methoxyestra-1,3,5 (10)-trien-17-one; 1amino-3-methoxy-19-norpregna-1, 3,5(10)-trien-20-one; 1amino-1lB-acetoxy-3-methoxyestra-l,3,5(10)-trien-17-one; l amino17,20;20,2l-bismethylenedioXy-3 methoxy 19 norpregna 1,3,5(10 trien-l-01 ll-acetate; 1 amino-3-methoxyestra 1,3,5

Patented Nov. 1, 1966 "ice (10)-trien-l6a,17fl-diol 16,17-diacetate;l-amino-B-methoXyestra-1,3,5 10)-trien-17fi-ol-16-one 17-acetate.

The l-amino-estratrienes described above on treatment with an alkalimetal nitrite under acid conditions at a low temperature yield diazoniumsalts which upon treatment with alkali metal azides will produce forexample 1-azido-3-methoxy-1,3,5(10)-estratrien-l7-one. The lattercompound on heating with hexadecane in an oxygenfree atmosphere willproduce 1,11-irnino-3-methoxyestra- 1,3,5(10) -trien-17-one. The lattermay also be prepared by irradiating with ultraviolet light a benzenesolution of the l-azido-compound. The hydrogen atom on the 1,1l-iminogroup may be replaced by a lower alkanoyl group by reaction with a loweralkanoyl chloride or an hydride, and by a lower alkyl group by reactionwith an alkyl halide in the presence of an alkali metal hydride.

The compounds directly convertible into the present compound may be forexample 1,1 1-imino-3 -methoxy-estra-1 ,3,5 10) -trien-17-one;

1,1l-imino-3-methoxyestra-1,3,5(10)-trien-17-one hydrochloride;

1,1 1-imino-3-hydroxyestra-l ,3,5 10) -trien-17-one;

1,1 1-propionylimino-3-methoxyestra-1,3 ,5 10) -tn'en- 17 one;

1,1 1-benzoylimino-3-methoxyestra-1,3,5 (10)-trien-17- one;

1,1 l-( 3',4',5'-trimethoxybenzoyl) imino-3 -methoxyestra-1,3,5(10)-trien-17-one;

1,1 1-imino-3-hydroxyestra-1,3,5(10)-trien-17-one hydrochloride;

1,1 1-imino-3 -hydroxyestra-1,3,5 10)-trien-17-one hydrosulfate;

1,1 1-acety1imino-3 -rnethoxyestra-l ,3,5 (10)-trien-17-one;

1,1 1-N-methylimino-3-methoxyestra-1,3,5 10) -trien-l7- one,

and the like.

By heating the above 1,11-irninoestratrienes with palladium-on-carboncatalyst the following 1,11-iminoestratetraenes of the present inventionare obtained:

and the like.

The 1,11-iminoestratetraenes of the present invention when administeredto rats produce a lowering of blood cholesterol and therefore are usefulas hypocholesterolemic agents. They also show bactericidal andfungicidal activity.

The following examples describe in detail the preparation ofrepresentative 1,1l-iminoestratetraenes of the present invention. 7 Q

Example 1.1-(p-nitrophenylazo) -3-mezh0xy-4-amin0- estra-1,3,5 (10-trien-1 7-0ne To a stirred solution of 7.0 g. (50.5 mmoles) ofp-nitroaniline in 50 m1. of glacial acetic acid and 75 ml. of 2.0 Nhydrochloric acid (150 mmoles) at -5 C. is added a solution of 3.48 g.(50.1 mmoles) of sodium nitrate in 15 ml. of water, below the surface ofthe liquid, and stirring is continued for 15 minutes. The resultingsolution of p-nitrobenzene diazonium chloride is poured into a wellstirred solution of 19.47 g. (50.0 mmoles) of 4-amino-3-methoxyestra-1,3,5 l)-trien 17 one (melting point 189-l93 C.) in250 ml. of glacial acetic acid and 25 ml. 5.0 N sodium hydroxide. (125mmoles) at room temperature. The deep red mixture is diluted with oneliter of water, allowed to stand overnight then filtered and the productwashed thoroughly on the filter with water. The yield of crude materialafter drying was 21.0 g. (94%), melting point 185l90 C.

The crude product, which contained some combined hydrogen chloride, isdissolved in methylene chloride (250 ml.), and then methanol (500 ml.)plus triethylamine (5.0 ml.) is added portionwise to the boilingsolution until all of the methylene chloride had been removed. Theresulting mixture gives 19.61 g. (87% yield) of very deep red crystals,melting point 242- 243 C. dec., which contains'only minor impurities bythin layer chromatographic analysis and is sufficiently pure for thenext step.

A pure sample is obtained by chromatography on activated magnesiumsilicate (60-100 mesh) using 15% ether-benzene as eluant.Crystallization of the product from methylene chloride-methanol, asabove, gives material of melting point 243 -244 C. dec.

Example 2 .1-(p-nitrophenylazo)-3-methoxyestra- 1,3,5 (10)-trien-1 7-0neTo a stirred solution of 17.94 g. (0.04 mole) of 1(pnitrophenylazo)-3methoxy 4 .aminoestra 1,3,5(10)- trien-17-one (melting point 242243dec.) in 400 ml. of glacial acetic acid and 100 ml. of 30% (w./w.)aqueous sulfuric acid at 0 C. is added a solution of 3.04 g. (0.044mole) of sodium nitrite in 30 ml. of water below the surface of theliquid. Stirring is continued at 0 C. for 15 minutes, then 450 ml. of50% aqueous hydrophosphorus acid is added and the mixture stirredovernight (16 hours) at room temperature. The precipitate is filtered,washed thoroughly with water and dried to give 16.4 g. (95% yield) ofcrude red-brown product, melting point 203 205 C. Crystallization frommethylene chloride-methanol gives 15.31 g. (89% yield) of red crystals,melting point 2l62l9 C. which contains only trace impurities by thinlayer chromatographic analysis and is pure enough for the next step.Chromatography of a sample on activated magnesium silicate (60-100mesh), using ether-benzene as eluant, followed by crystallization of theproduct from methylene chloridemethanol gives an analytical sample,melting point 223-224 C.

Example 3.I-amin0-3-methoxyestra-1,3,5 ()-trien- 17-0ne A solution of13.0 g. (0.03 mole) of l-(p-nitrophenylazo)-3methoxyestra-1,3,5(10)trien 17 one (melting point 215 217 C.) in 100 ml. of methylene chlorideis added to a stirred mixture of 30 g. of zinc dust in 300 ml. ofglacial acetic acid over approximately 10 minutes. The initialtemperature of 23 soon rises to 40-45 C. and is maintained in this rangeduring the reaction by occasional cooling in a water bath. An additional30 g. of zinc dust is added after half of the steroid is fed in. Themixture is stirred for a further 10 minutes then filtered and theresidue of zinc washed on the filter with acetic acid. The filtrate isconcentrated under reduced pressure to approximately 125 ml, dilutedwith 500 ml. of water and extracted with chloroform. The extract iswashed fate, concentrated to a volume of 100 ml. and filtered through abed of magnesium silicate (60 g.) using 300 ml. of chloroform wash. Thefiltrate is evaporated under reduced pressure to a small volume andcrystallized from methanol to give 5.58 g. of grey solid melting point198208 C. which contains a trace of .p-phenylenedie amine impurity. Anadditional 1.0 g. of product, melting point 209213 C. is obtained bychromatography of the filtrate on activated magnesium silicate 60-100mesh) using 20% ethyl acetate-n-hexane as eluant, followed bycrystallization of the product from methylene chlorideether. The totalyield of product is therefore 6.58 g. (73%).

Analytically pure material is obtained by chromatog raphy of a sample onactivated magnesium silicate as above, followed by crystallization frommethanol to give a white crystalline product, melting point 213-214 C.

Example 4.1-amin0-3-meth0xyestra-1,3,5 (10 trien-1 75-01 Sodiumborohydride (1.9 g.) is added ,to a suspension of1-amino-3-methoxyestra-1,3,5( 10)-trien 17 one (3.0 g.) in methanol (200ml.). The resulting mixture is stirred for 30 minutes at roomtemperature, acidified with acetic acid and evaporated. The residue ispartitioned between ether and water, and the ether phase is dried overanhydrous sodium sulfate and evaporated to give the product of theexample.

Example 5.1-azid0-3-methoxyestra-1,3,5 (10 -trien- 17-0ne To a stirredsolution of 14.0 g. (0.046 mole) of 1- amino-3-methoxyestra-1,3,5 l0)trien 17 one (melting point 198-208 C.) in 200 ml. of acetic acid plus139.8 ml. (0.139 equivalent) of 1.0 N sulfuric acid at 20 C. (Dry Iceacetone bath) is added a solution of 3.55 g. (0.051 mole of sodiumnitrite in 10 m1. of water below the surface of the liquid. The solutionis stirred for 2 minutes then a solution of 12.1 (0.186 mole) of sodiumazide in 50 ml. of water is added as quickly as the resulting vigorousevolution of nitrogen would allow. The orange col ored mixture isstirred at 10 to +5 C. for 30 minutes then extracted with chloroform andthe extract washed with water, saturated sodium bicarbonate and finallywith water. After drying over magnesium sulfate, the solution isevaporated under reduced pressure to an oil which crystallizes to a darkbrown solid on cooling. The total crude product. is chromatographed on500g. of activated magnesium silicate (60-100 mesh) using 7.5% ethylacetate-n-hexane as eluant. The yield of cream colored solid is 12.1 g.melting point 144-148 C. A single crystallization of a sample frommethylene chloride-methanol gives analytical material with melting point147 149 C.

Example 6.1-azido-3-methoxy-1,3,5 (1 0) -estratrien-1 7,8-01

Following the procedure of- Example 5 1-amin0-3-methoxyestra-1,3,5(10)-trien-17;3-0l is converted into the product ofthe example.

Example 7.1,11-imino-3-metlz0xyestra-1,3,5(10)- trien-1 7 -one Solid1-azido-3-methoxyestra-1,3,5 (10) trien 17-one (10 g., 0.031 mole,melting point 144-148 C.) is added in one portion with stirring tohexadecane at 200 C. in an atmosphere of argon and'heating continued fora total of five minutes. The hexadecane is previously purified byshaking with several portions of concentrated sulfuric acid followed bywashing with water, drying over magnesium-sulfate and storing overmolecular sieves. The reaction mixture is allowed to stand overnight atroom temperature. Filtration gives 7.92 g. (86% yield) of crude productwhich is chromatographed on magnesium silicate (6.00 g. of 60-100 mesh).Elution with 15% and finally 20% ethyl acetate-petroleum ether (SO-75)gives 7.2 g. of product which still contains a less polar impurity. Thisis removed by dissolving the product in 250 ml. of 0.74 N hydrochloricacid and extracting the solution with ether. The product is recovered bymaking the aqueous phase basic with excess 5.0 N sodium hydroxide andextracting the precipitate with methylene chloride. Evaporation of themethylene chloride'extract under reduced pressure'gives an oil. whichcrystallizes on trituration with ether giving 6.7 g. of cream coloredproduct, melting point 192198 C. Crystallization from methanol gives5.94 g. (65% yield) of white product melting point 199201 C. which ispure by thin layer chromatographic analysis.

An additional crystallization of a sample from methanol did not alterthe melting point of 199201 C.

The 1,11-imino-compound is also prepared by irradiating with an Hanoviahigh pressure ultraviolet lamp a benzene solution of thel-azido-compound.

Example 8.1,11-imin-3-meth0xyestra-1,3,5 (1 0) trien-l 7B-ol Followingthe procedures of Example 7 1-azido-3- methoxyestra-1,3,5()-t1ien-17B-ol gives the product of the example.

Example 9.1,11-imin0-3-methoxyestra-1,3,5 (10 9 (11 )-tetraen-1 7 -one Amixture of 892.1 mg. (3.0 millimoles) of 1,11-immo-3-methoxyestra-l,3,5(10)-trien 17 one melting point 199-201 C. and 300mg. of 10% palladium-on-carbon in 45 ml. of xylene is stirred andrefluxed for one hour. After cooling to room temperature, the mixture isfiltered through 18 g. of magnesium silicate which is washed withn-hexane to remove the xylene and the product dissolve-d with 250 ml. ofmethylene chloride. Evaporation of the methylene chloride filtrate gives846.6 mg. (95% yield) of white crystalline product, melting point 197201" C. Crystallization from methanol in the presence of one drop ofpyridine gives 738.5 mg, melting point 202-204 C., plus a second crop of56 mg., melting point 199 203 C. Hence, a total yield of crystallizedproduct is 794 mg. (90%). A single recrystallization from methanol givesan analytically pure sample, melting point 203 205 C.

Example 1 0.-1,11-imino-3-metlzoxyestra-1,3,5 (10) 9 (11 )-tetraen-176-01 Following the procedure of Example 9 1,11-3-methoxyestra-1,3,5(10)-trien-17fi-ol gives the product of the example.

Example 11 .1,11-imin0-3-meth0xyestra-1,3,5 (10 9 (11 -tetraen-1 7onehydrochloride Example 12.1,11-imin0-3-hydr0xyestra-1,3, (10

9 (11 )-tetraen-I 7-one Treatment of 1,11-imin0 3 methoxyestra-1,3,5(10) 9(l1)-tetraen-l7-one with pyridine hydrochloride gives the productof the example.

Example 13.-1,11-imin0estra-1,3,5(10) ,9(11 tetraene-3,1 7p-di0lTreatment of 1,11-imino 3 methoxyestra-1,3,5(10), 9(11)-tetraen-175-o1with pyridine hydrochloride gives the product of the example.

Example 14.1,11-acetylimino-3-methoxyestra-1,3,5-

(10) ,9 (11 -tetraen-17-one To a solution of 0.5 g. of1,11-imino-3-methoxyestra- 1,3,5(10),9(11)-tetraen-17-one in 10 ml. ofpyridine is added 10 ml. of acetic anhydride. The mixture is heated on asteam bath for 1 hour and then allowed to stand at room temperatureovernight. The excess acetic anhydride is decomposed with methanol andthe solution is evaporated. The residue is partitioned betweenethermethylene chloride (2:1) and 5% sodium carbonate solution. Theorganic phase is dried and evaporated to give the product of theexample.

Example 15.I,11-propionylimin'o-3-methoxyestra- 1,3,5 (10 ,9 (11)-tetraen-1 7 one Treatment of 1,11-imino 3 methoxyestra-1,3,5 (10),9(11)-tetraen-17-one in pyridine with propionic anhydn'de on the steambath for 1 hour followed by standing overnight at room temperature givesthe product of the example.

Example 16.1,11-benz0ylimino-3-methoxyestra-1,3,5(10),9(11)-tetraen-17-0ne Substitution of benzoyl chloride forpropionic anhydride in Example 15 gives the product of the example.

Example 1 7.--1,11- (3,4',5'-trimeth0xybenz0yl) iminoestra-1,3,5 (10),9(11 -tetraen-1 7-0ne Substitution of 3,4,5-trimethoxybenzoyl chloridefor propionic anhydride in Example 15 gives the product of the example.

Example -I8.I,11-N-methylimin0-3-meth0xyestra- 1,3,5 (10) ,9 (11-tetraen-17-one A solution of 1,11-imino-3-methoxyestra 1,3,5(10), 9( 11 -tetraen-17-one in 20% dimethylformamide-benzene is treated withsodium hydride and methyl iodide. The mixture .is heated at 40 C. for 1hour, cooled and extracted with benzene. The extract is washed withwater and evaporated to give the N-met-hyl derivative, the product ofthe example.

Example 19.1,11-N-methylimino-3-methoxyestra 1,3,5 (1 0),9(11)-tetraen-1 7-0ne methiodide A solution of1,11-N-methylimino-3-methoxyestra-1,3, 5(10),9(11) tetraen-17-one inether-methanol is treated with methyl iodide to give the methiodide.

Example 20.-1,11-imin0-3-hydroxyestra-1,3,5(10), 9 (11 -tetraen-1 7 -onehydrochloride Treatment of 1,11 imino-3-hydroxyestra-1,3,5(10),9(11)-tetraen-17-one in a mixture of ether and hydrogen chloride givesthe product of the example.

Example 21 .1,11-imino-3-methoxyestra-1,3,5(10), 9 (11 tetraen-17-0nehydrosulfate Treatment of 1,11 imino-3-hydroxyestra 1,3,5(10),9(11)-tetraen-17-one in a mixture of ether and methanol with sulfuricacid gives the product of the example.

Example 22.-1,11-imin0-17a-ethynylestra-1,3,5 (10) 9 (11 )-zetraen'e-3,1 7 fl-di ol A solution of potassium acetylide in liquidammonia is prepared by bubbling acetylene into a solution containing 3g. of potassium in ml. of liquid ammonia until the blue colordisappears. To it is added dropwise a solution of 1.3 g. of1,1l-imino-3-hydroxyestra-1,3,5(10), 9(1'1)-tetraen-17-one in 75 m1. ofether. After 2 hours the cooling bath is removed and the mixture isallowed to warm to room temperature and stored overnight. Dilutesulfuric acid (8%, 70 ml.) is added dropwise to the mixture and it isextracted with ether. The extract is washed with saturated sodiumchloride solution, dried and evaporated .to give the product of theexample.

7 Example 23 .-1 ,1 1 -imin-l 7a-vinylestra-1,3,5 9 (11 )-tetraene-3,1 7fl-diol Example 24.1,11-imin0-1 7a-methylestra-1,3,5(1 0 9 (I 1)-tetraene-3,1 7 ,B-diol To a freshly prepared solution of methylmagnesium iodide (4.2 ml. of methyl iodide and 1.6 g. of magnesiumturnings) in ether (30 ml.) is added a solution of 1,11-imino-3-hydroxyestra-1,3,5 10 ,9 1 1 -tetraen-17-one (0.9 g.) in ether(25 ml).

wise addition of excess ammonium chloride solution. The ether phase isseparated and the aqueous phase is extracted several times withmethylene chloride. The combined extracts are washed with water, driedand evaporated to give the product of the example.

We claim:

1. A compound of the formula:

wherein R is selected from the group consisting of hydrogen and loweralkyl; Rv is selected from the group consisting of hydrogen, loweralkyl, lower alkanoyl, mono-nuclear aroyl, and lower alkoxy mono-nucleararoyl; X is selected from the group consisting of -loweralkyl 011:011,

The resulting mixture is heated. to reflux for 6 hours, cooled andquenched by the dropand / czorr and acid addition and quaternaryammonium salts thereof.

2. The compound 1,11 imino 3 methoxyestra-1,3,5(10),9(11)-tetraen-17-one.

3. The compound 1,11 imino 3 methoxyestra-1,3, 5 10) ,9 1 1)-tetraen-17-one hydrochloride.

4. The compound 1,11 imino 3 hydroxyestra-1,3, 5(10),9(11)-tetraen-17-one.

5. The compound 1,11 imino 3 hydroxyestra-1,3, 5 (10),9(11)-tetracn-17-one hydrochloride.

6. The compound 1,11 imino 3 hydroxyestra-1,3, 5 10) ,9 1 1-tetraen-l7-one hydrosulfate.

7. The compound 1,11 imino 3 methoxyestra1,3, 5(10),9( ll)-tetraen-17fl-ol.

8. The compound 1,11 iminoestra 1,3,5(10),9(11)- tetraene-3,17,B-diol.

9. The compound 1,11 acetylimino 3 methoxyestra- 1,3,5 10) ,9 1 1)-tetraen-l7-one.

10. The compound 1,11 propionylamino 3methoxyestra-1,3,5(10),9(11)-tetraen-17-one.

11. The compound 1,11 benzoylimino 3 methoxy 16. The compound 1,11 imino17o: vinylestra-1,3,

5 l0),9( l l )-tetraene-3,17fl-diol.

17. The compound 1,11 imino 17oz methylestra-1,3, 5 l0) ,9 l 1)-tetraene3,17 8-diol.

No references cited.

LEWIS GOTTS, Primary Examiner.

E. L. ROBERTS, Assistant Examiner.

1. A COMPOUNDD OF THE FORMULA:1,11-(-N(-R'')-),3-(R-O-)ESTRA-1,3,5(10),9(11)-TETRAENE WHEREIN R ISSELECTED FROM THE GROUP CONSISTING OF HYDROGEN AND LOWER ALKYL; R'' ISSELECTED FROM THE GROUP CONSISTING OF HYDROGEN, LOWER ALKYL, LOWERALKANOYL, MONO-NUCLEAR AROYL, AND LOWER ALKOXY MONO-NUCLEAR AROYL; X ISSELECTED FROM THE GROUP CONSISTING OF -CO-; -CH(-OH)-; -C(-OH)(-LOWERALKYL)-; WHERE CARBON 17 OF RING IS REPLACED WITH X -C(-OH)(-CH=CH2)-AND -C(-OH)(-C*CH)AND ACID ADDITION AND QUATERNARY AMMONIUM SATSTHEREOF.